Differential diagnosis of non-molar gestational trophoblastic neoplasia with ectopic pregnancy by clinical-pathological features.

PURPOSE: This study was presented to investigate the clinical-pathological characteristics of gestational trophoblastic neoplasia (GTN) following non-molar pregnancy and differentiated with ectopic pregnancy (EP). METHODS: The clinical data of 83 patients who were admitted for suspected GTN after non-molar pregnancy at the Women's Hospital School of Medicine Zhejiang University from January 2015 to September 2022 were selected for analysis. RESULTS: In total, 41 cases were confirmed non-molar GTN, including 31 choriocarcinoma, 9 PSTT (placental site trophoblastic tumor), and 1 ETT (epithelioid trophoblastic tumor), while 42 cases were confirmed EP. Compared with ectopic pregnancy, non-molar GTN patients had lower levels of serum progesterone compared with EP (3.81 nmol/L vs 17.70 nmol/L, P = 0.001). Based on the ultrasound, the thickness of the endometrium was thinner in patients with non-molar GTN compared with EP (0.565 cm vs 0.70 cm, P = 0.018). By histopathologic examination, the endothelium of non-molar GTN showed less decidual-like changes compared with EP (64.3% vs 14.6%, P = 0.001). CONCLUSION: A combination of serum progesterone levels, endometrium thickness, and histopathologic features of the endometrium can help to differentiate non-molar GTN and EP. Surgeries including hysteroscopy with curettage and/or laparoscopy are needed.

Squamous cell carcinoma of the cervix associated with choriocarcinomatous differentiation: a case report and review of the literature.

The morphology of the choriocarcinomatous variant of cervical squamous cell carcinoma (SCC) suggests an undifferentiated aggressive biological behaviour and a poor outcome, for which standard treatment has not been established. In addition, cases are rarely reported, with only five cases of patients with cervical carcinoma with choriocarcinoma reported previously in the literature. This current case report describes in detail a patient who was diagnosed with cervical SCC mixed with choriocarcinomatous differentiation. The case report includes details of the diagnosis, pathology, short tandem repeat genotyping, treatment and follow-up of this patient. As there is no standard treatment for this variant, the patient underwent surgery followed by radiotherapy. Unfortunately, 4 months after therapy discontinuation, radiological evaluation and laboratory tests documented a recurrence of the disease and the patient died. This report also systematically reviews the literature on cervical cancer associated with choriocarcinomatous differentiation and the five previous cases. It provides the most up-to-date summary of this disease, including its clinical manifestations, histopathology, diagnosis, treatment and prognosis.

MicroRNA profile of paclitaxel-resistant serous ovarian carcinoma based on formalin-fixed paraffin-embedded samples.

BACKGROUND: To assess the feasibility of validating microRNA (miRNA) profile related to paclitaxel-sensitivity in formalin-fixed paraffin-embedded (FFPE) samples of serous ovarian carcinoma (OC) patients. METHODS: Deregulated miRNAs identified by miRNA microarray were further detected in 45 FFPE OC samples using Realtime PCR. Correlations between paired FFPE and frozen tumor samples were analyzed. Survival times were compared between 6 high and low miRNAs groups. Western blot and luciferase reporter assay were used for validating the target of miRNA. RESULTS: Sixteen up-regulated miRNAs and twenty-three down-regulated miRNAs were revealed in pacilitaxel-resistant ST30 cells. The up-regulated miRNAs (miR-320a, 22 and 129-5p) and down-regulated miRNAs (miR-9, 155 and 640) were confirmed in paclitaxel-resistant FFPE tumor samples, compared with paclitaxel-sensitive samples. Higher miR-9 and miR-640 showed better survival time in OC patients. Expressions of miR-9, 155 and 22 in FFPE samples were closely mimicked by those in frozen tissues. RAB34 was validated as a direct target of miR-9. CONCLUSIONS: We validated miRNA profile in pacilitaxel-resistant OC using FFPE samples, which might enable treatment stratification and help us to predict outcomes in OC patients. FFPE samples are feasible materials for miRNA research.

Case report: Squamous cell carcinoma and spindle cell sarcoma (SCS) arising in a mature cystic teratoma of the ovary.

Objective: Malignant transformation of mature ovarian teratoma is a rare phenomenon, mainly occurring in postmenopausal period. Squamous cell carcinoma accounts for 80% of all malignant transformations. Sarcoma transformation is much less common and tends to imply a poorer prognosis and aggressiveness. Case report: We report a case of undifferentiated sarcoma with squamous cell carcinoma in a mature cystic teratoma of the ovary in a 36-year-old woman. The tumor shows epithelial and stromal components. This is a unique report of a benign teratoma of the ovary with malignant transformation, showing epithelial and sarcomatous components. This young woman presented with abdominal distension and a rapidly enlarging ovario-derived pelvic mass with a slightly elevated CA199 tumor marker of 115.9 U/ml. The woman underwent transabdominal excision of the left ovarian cyst on October 20, 2020. During the operation, rapid freezing pathological examination did not indicate malignancy. The postoperative paraffin pathology revealed undifferentiated sarcoma with squamous cell carcinoma (from mature cystic teratoma malignancy), and she finally received comprehensive staging surgery. Postoperative paraffin pathology showed no residual cancer in uterus and other tissues, and all lymph nodes were negative. The patient was finally diagnosed with ovarian malignant tumor IC1 stage (high-grade spindle cell sarcoma complicated with squamous cell carcinoma). Chemotherapy was completed three times after surgery, and no signs of recurrence were found after follow-up. Conclusion: The preoperative diagnosis and intraoperative rapid freezing examination of malignant transformation of mature teratoma of ovary are challenging.

Molecular profile-based recommendations for postoperative adjuvant therapy in early endometrial cancer with high-intermediate or intermediate risk: a Chinese randomized phase III trial (PROBEAT).

BACKGROUND: The use of molecular categorisation is shifting paradigm towards the use of molecular information to refine risk stratification in endometrial cancer (EC). To date, evidence to support molecular-guided therapies is limited to retrospective studies and secondary molecular analyses of patients receiving standard treatment. The PROBEAT study is the first randomized phase III trial to evaluate tailored adjuvant treatment based on WHO-endorsed molecular classification in Chinese EC patients. It is expected to provide a clinical decision-making tool for adjuvant treatment of patients with high-intermediate risk (HIR) or intermediate risk (IR) EC to better optimise and personalise patient care and increase relapse-free survival. METHODS: The PROBEAT trial is a prospective, multicentre study led by Women's Hospital of Zhejiang University Gynaecologic Oncology Group. Recruitment started on January 24, 2022, and 590 patients with HIR or IR endometrioid EC are expected to be recruited from 13 clinical centres in China. All tumor tissues will be classified into four molecular subtypes (POLEmut, MMRd, p53abn, or NSMP) based on WHO-endorsed molecular classification. Patients will be randomly assigned at a 2:1 ratio to either experimental arm and will receive molecular profile-based adjuvant treatment (observation in the POLEmut subgroup, vaginal brachytherapy in the MMRd or NSMP subgroup, or chemoradiotherapy in the p53abn subgroup) or to standard arm and will receive preferred adjuvant radiotherapy as recommended by the recent National Comprehensive Cancer Network guidelines version 1 (2022). The primary outcome is 3-year rates of recurrence. Secondary outcomes are relapse-free survival, overall survival, adverse events and health-related cancer-specific quality of life. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT05179447.

Adjuvant chemotherapy versus adjuvant concurrent chemoradiotherapy after radical surgery for early-stage cervical cancer: a randomized, non-inferiority, multicenter trial.

We conducted a prospective study to assess the non-inferiority of adjuvant chemotherapy alone versus adjuvant concurrent chemoradiotherapy (CCRT) as an alternative strategy for patients with early-stage (FIGO 2009 stage IB-IIA) cervical cancer having risk factors after surgery. The condition was assessed in terms of prognosis, adverse effects, and quality of life. This randomized trial involved nine centers across China. Eligible patients were randomized to receive adjuvant chemotherapy or CCRT after surgery. The primary end-point was progression-free survival (PFS). From December 2012 to December 2014, 337 patients were subjected to randomization. Final analysis included 329 patients, including 165 in the adjuvant chemotherapy group and 164 in the adjuvant CCRT group. The median follow-up was 72.1 months. The three-year PFS rates were both 91.9%, and the five-year OS was 90.6% versus 90.0% in adjuvant chemotherapy and CCRT groups, respectively. No significant differences were observed in the PFS or OS between groups. The adjusted HR for PFS was 0.854 (95% confidence interval 0.415-1.757; P = 0.667) favoring adjuvant chemotherapy, excluding the predefined non-inferiority boundary of 1.9. The chemotherapy group showed a tendency toward good quality of life. In comparison with post-operative adjuvant CCRT, adjuvant chemotherapy treatment showed non-inferior efficacy in patients with early-stage cervical cancer having pathological risk factors. Adjuvant chemotherapy alone is a favorable alternative post-operative treatment.

Chemotherapy initiation with single-course methotrexate alone or combined with dactinomycin versus multi-course methotrexate for low-risk gestational trophoblastic neoplasia: a multi-centric randomized clinical trial.

We aimed to evaluate the effectiveness and safety of single-course initial regimens in patients with low-risk gestational trophoblastic neoplasia (GTN). In this trial (NCT01823315), 276 patients were analyzed. Patients were allocated to three initiated regimens: single-course methotrexate (MTX), single-course MTX + dactinomycin (ACTD), and multi-course MTX (control arm). The primary endpoint was the complete remission (CR) rate by initial drug(s). The primary CR rate was 64.4% with multi-course MTX in the control arm. For the single-course MTX arm, the CR rate was 35.8% by one course; it increased to 59.3% after subsequent multi-course MTX, with non-inferiority to the control (difference -5.1%,95% confidence interval (CI) -19.4% to 9.2%, P = 0.014). After further treatment with multi-course ACTD, the CR rate (93.3%) was similar to that of the control (95.2%, P = 0.577). For the single-course MTX + ACTD arm, the CR rate was 46.7% by one course, which increased to 89.1% after subsequent multi-course, with non-inferiority (difference 24.7%, 95% CI 12.8%-36.6%, P < 0.001) to the control. It was similar to the CR rate by MTX and further ACTD in the control arm (89.1% vs. 95.2%, P =0.135). Four patients experienced recurrence, with no death, during the 2-year follow-up. We demonstrated that chemotherapy initiation with single-course MTX may be an alternative regimen for patients with low-risk GTN.

Management and prognostic analysis of patients with gestational trophoblastic neoplasia (GTN) in FIGO stage IV and its special type.

GTN is a group malignant diseases from placental trophoblastic cells. There are very few cases of GTN with FIGO (International Federation of Gynecology and Obstetrics) stage IV all over the world, and the special types (patients with metastatic lesions and with no evidence of GTN neither in genitalia nor in lungs) have rarely been reported. It is necessary to conduct large retrospective studies aimed at exploring the diagnosis, treatment and outcomes of this disease. In this retrospective study, 716 patients with GTN were treated at Zhejiang University School of Medicine Women's Hospital between January 1999 and September 2019; 26 patients were diagnosed as stage IV GTN; Among the 26 stage IV GTN patients, 5 were defined as the special types. The 5-year OS rate of the total 26 FIGO stage IV GTN patients was 69.0%. There was no significant difference of survival rate between stage IV GTN and its special type. And no significant differences in blood type, antecedent pregnancy type, the interval from last known pregnancy, pretreatment serum HCG (human chorionic gonadotropin) level, maximum diameter of tumors, FIGO score, underwent surgery or not and pathological pattern by the outcomes. Age, number of tumor lesions, primary chemotherapy regimen was EMA-CO or EP-EMA protocol and chemoresponse affected the prognosis significantly. Only number of tumor lesions > 8 was independent prognostic factors associated with poorer OS.

[Study on high risk factors associated with positive margin of cervix conization in patient with cervical intraepithelial neoplasia].

OBJECTIVE: To assess the high risk factors associated with the positive margin of conization in patients with cervical intraepithelial neoplasia (CIN). METHODS: From January 2000 to February 2008, 1699 consecutive patients with CIN undergoing conization was reviewed retrospectively in order to analyze the relationship between the positive margin of conization with clinical prognostic factors, such as patients age, disease grade, size of lesion, the procedure of excision and menopause. chi2 tests was used to compare the different frequencies of factors in groups of positive and negative margin conization, then seven factors with positive margin were processed into unconditional logistic regression analysis. RESULTS: The rate of the positive margin in 1699 patients was 14.01% (238/1699). The mean age of patients with positive margins was (39+/-9) years old, while patients with negative margins was (39+/-8) years old, which didn't reach statistical difference (P>0.05). The rate of the positive margin was 8.63% in cold knife cone (CKC) and 18.66% in loop electrosurgical excision procedure (LEEP), which showed significant difference (P<0.01). Among 1699 patients, 90 patients were with CINI, 339 patients were with CIN II, 1113 patients were with CIN III [including 972 with severe dysplasia and 141 with cancer in situ (CIS)], 87 patients were with cervical cancer stage Ia1, 70 patients were with stage Ia2 or advanced stages. The rate of positive margin was 1.11% (1/90), 3.83% (13/339), 10.70% (104/972), 26.24% (37/141), 35.63% (31/87) and 74.29% (52/70), respectively. There was statistic difference among them, except CINI and CINII. When combined CIN I with CIN II, then compared with CIN III, cervical cancer with Ia1 and Ia2, it also showed statistical difference (P<0.05). The rate of positive margin in postmenopausal women was 21.54% (28/130), which was significantly higher than 13.38% (210/1569) in premenopausal women (P=0.010). The logistic regression analysis showed that the procedure of excision, grade of disease, size of lesion, surface of cervix, and menopause were high risk factors associated with the positive margin, the risk ratio were 5.147, 3.048, 1.271, 1.905 and 1.860, respectively. CONCLUSIONS: High grade, the extent of CIN disease, LEEP and postmenopausal age are high-risk factors associated with positive margin in patients treated by conization. It should be warranted in those patients when designing conization treatment.

Whole-exome sequencing reveals genetic variants in ERC1 and KCNG4 associated with complete hydatidiform mole in Chinese Han women.

Complete hydatidiform mole (CHM) is a rare pregnancy-related disease with invasive potential. The genetics underlying the sporadic form of CHM have not been addressed previously, but maternal genetic variants may be involved in biparental CHM. We performed whole-exome sequencing of 51 patients with CHM and 47 healthy women to identify genetic variants associated with CHM. In addition, candidate variants were analyzed using single base extension and Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry in 199 CHM patients and 400 healthy controls. We validated candidate variants using Sanger sequencing in 250 cases and 652 controls, including 205 new controls. Two single nucleotide polymorphisms, c.G48C(p.Q16H) inERC1 and c.G1114A(p.G372S) in KCNG4, were associated with an increased risk of CHM (p<0.05). These variants may contribute to the pathogenesis of CHM and could be used to screen pregnant women for this genetic abnormality.

[Effects of primary chemotherapy with single methotrexate on low-risk gestational trophoblastic neoplasia and influencing factors thereof].

OBJECTIVE: To evaluate the effects of primary chemotherapy with single-agent methotrexate (MTX) on low-risk gestational trophoblastic neoplasia (GTN) and the influencing factors thereof. METHODS: Sixty-one GTN patients with the score of < or = 6 according to the new International Federation of Gynecology and Obstetrics (FIGO) scoring system (2000) were divided into 2 groups: 51 patients were treated with single MTX 0.4 mg/kg daily for 5 days (MTX 5 d group), and 10 patients were treated with MTX on the days 1, 3, 5, and 7, and with folinic acid (FA) 0.1 mg/kg on the days 2, 4, 6, and 8 (MTX + FA group), both group with an interval of treatment course of 2 weeks. The serum level of human chorionic gonadotropin (hCG) was detected every week. If a plateau or increase of serum hCG appeared between 2 examination results, meaning tolerance to MTX, the patients concerned had to undergo different regimens of salvage chemotherapy, all with MTX as one of their components. Univariate and multivariate methods were used to analyze the relationships of different factors to the outcomes of chemotherapy. RESULTS: Thirty-five of the 51 patients of the MTX 5d group (68.6%) achieved complete primary remission, 3 of the 10 patients of the MTX + FA group achieved complete primary remission, and all patients achieved complete remission after salvage chemotherapy. Univariate analysis showed that the mean pretreatment serum level of hCG, duration between antecedent pregnancy and start of treatment, size of tumor, FIGO score, specific regimen of MTX were significantly associated with outcome of chemotherapy (P = 0.004, 0.022, 0.017, 0.005, 0.021 respectively). Logistic regression analysis showed that only three independent factors predictive for the outcome of chemotherapy: MTX regimen (OR = 2.476), FIGO score (OR = 1.431), and pretreatment hCG titer (OR = 1.001). CONCLUSION: Primary chemotherapy with single MTX regimen may still be one of the options for patients with low-risk GTN according to the new FIGO scoring system, though the rate of complete primary remission appears to be lower. All patients with low-risk GTN achieve complete remission after salvage chemotherapy. MTX regimen, FIGO score, and pretreatment hCG are independent risk factors of outcome of single MTX chemotherapy.

[Efficacy and side effects of methotrexate in the treatment of ectopic pregnancy].

OBJECTIVE: To study the efficacy and side effects of methotrexate with different protocols in the treatment of ectopic pregnancy (EP). METHODS: Total of 648 patients who had EP and were treated only with MTX were analysed. Patients were divided into six groups according to protocol: Group 1, 100 mg intravenously (IV); Group 2, 100 mg, IV, followed by citrovorum factor; Group 3, 20 mg, IV every day for five days; Group 4, 20 mg intramusculary (IM) every day for five days; Group 5, 75 mg, IV; Group 6, 75 mg, IM. RESULTS: The rates of repeated MTX injection in group 1 - 6 because of inadequate decrease of hCG were 23.3%, 25.0%, 21.4%, 20.6%, 24.4% and 22.4% respectively. Success rates were 87.4%, 85.4%, 90.5%, 92.6%, 86.3% and 91.4% respectively. Rates of liver dysfunction were 10.3%, 8.3%, 64.3%, 69.1%, 8.7% and 31.0%. CONCLUSION: Single-dose of 75 mg MTX IV injection may be the best regimen in the treatment of EP because of the same efficacy but the least side effects.

[cDNA microarray analysis of gene expression in acquired methotrexate-resistant of human choriocarcinoma].

OBJECTIVE: To identify the molecular components involved in acquired methotrexate-resistance of human choriocarcinoma. METHODS: Methotrexate-resistant cell line (JAR/MTX) was derived from JAR cell line by exposed to intervally and progressively increasing higher concentration of MTX. cDNA microarray analyses of JAR/MTX and its parental cell line JAR were performed. RESULTS: JAR/MTX was established after one year with stable resistance. Its resistant index to MTX was 7.3. Nine genes were differential expressed between JAR/MTX and JAR cells. INSR, SLC1A3, SAT, HBB, and FLJ12443 were underexpressed and HS1, TXNRD1, TAGLN2, and EEF2 were overexpressed in JAR/MTX cells. CONCLUSION: The cDNA microarray system showed that several alterations of gene expression were present in acquired methotrexate-resistance of human choriocarcinoma.

[Establishment of methotrexate-resistant human choriocarcinoma cell line and its biological characteristics].

OBJECTIVE: To establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics. METHODS: MTX-resistant cell line (JAR/MTX) was derived from human choriocarcinoma cell line JAR by exposed to intermittently and progressively increasing concentration of MTX. Drug sensitivity was detected by MTT; P-gp GST-Pi and PCNA expressions were detected by immunohistochemistry. Cell apoptosis was detected by flow cytometry (FCM) with PI/Annexin V stain. Growth rates and human chorionic gonadotropin (HCG) production were also measured. RESULTS: JAR/MTX cell line was established with stable MTX-resistance (resistance index to MTX was 7.3) and cross-resistant to TAX and VCR. Growthrate of JAR/MTX was lower than that of parent cell line JAR. Expression level of PCNA in JAR/MTX was lower than that in JAR (3.09+/-0.42 compared with 3.72+/-0.35, P<0.05), while GST-pi expression was higher. No statistical difference of P-gp expression existed between two cell lines. JAR/MTX secreted more HCG than JAR every 10(5) cells secreted (95.7+/-5.4 compared with 41.3+/-2.8)mIU after 48 h(P<0.01). The flow cytometry showed that the spontaneous and MTX induced apoptosis in JAR/MTX was significantly lower than that in JAR P<0.05. CONCLUSION: JAR/MTX cell line presented stable resistant to MTX and cross-resistant to TAX and VCR, which might sever as a model in study of drug resistance in choriocarcinoma.

MicroRNA detection in cervical exfoliated cells as a triage for human papillomavirus-positive women.

BACKGROUND: Papanicolaou (Pap) triage, with high specificity, has been recommended for primary Human papillomavirus (HPV) testing but is flawed by poor sensitivity and cytologist dependence. We evaluated the potential role of microRNA (miRNA) detection in cervical exfoliated cells in HPV-positive women from a clinic-based population. METHODS: Primary HPV testing as well as Pap test were performed on all eligible women. Six miRNAs (miR-424/miR-375/miR-34a/miR-218/miR-92a/miR-93) were detected by RT-qPCR in cervical exfoliated cells. All HPV-positive women underwent colposcopy and further biopsy if indicated. Mann-Whitney U test, the receiver operating characteristic curve, logistic regression, and Pearson's Chi-square were used to assess data. All tests of statistical significance were two-sided. RESULTS: A total of 1021 eligible HPV-positive women were enrolled. The expression of miR-424/miR-375/miR-34a/miR-218 in high-grade cervical intraepithelial neoplasia (CIN) and abnormal cytology was statistically significantly lower than that in low-grade CIN and normal cytology, respectively (all P < .05). Compared with the Pap test, both miR-424 and miR-375 detection achieved higher sensitivity (76.0% and 74.9% vs 63.8%, P < .05), higher negative predictive value (NPV) (85.7% and 85.4% vs 79.3%, P < .05), and comparable specificity while identifying CIN2 or worse (CIN2+). Similar results were achieved while identifying CIN3+. Multi-marker panels based on miR-424, miR-375, and miR-218 further improved the performance over any single miRNA test or Pap test. CONCLUSION: Single miR-424 or miR-375 detection and miR-424/miR-375/miR-218-based multimarker panels in cervical exfoliated cells show superior performance over Pap triage for high-grade CIN identification in a clinic-based population. Detection of miRNA may provide a new triage option for HPV-positive women.

FTY720 and cyclosporin protect ovarian tissue grafted into rabbits.

OBJECTIVE: To determine whether FTY720 combined with CsA has immunomodulatory effects on human ovarian tissue transplanted to the back muscle of rabbits for an 8-week period. STUDY DESIGN: We selected rabbits as recipients of ovarian xenografts with and without treatment by CsA and FTY720. Ovarian fragments from twelve patients were cut into 2 mm × 2 mm, 1-2mm thick pieces and randomly distributed into four groups: Group 1 (FTY720 2 mg/kg/d+CsA 3 mg/kg/d), Group 2 (FTY720 1 mg/kg/d+CsA 3mg/kg/d), Group 3 (FTY720 0.5 mg/kg/d+CsA 3mg/kg/d) and Group 4 for control (CsA 3 mg/kg/d). FTY720 was started three days before transplantation and was given daily after transplantation. CsA was administrated post-transplantation. All the animals were killed 8 weeks post- transplantation. Levels of serum estrogen (E2), interferon-γ (IFN-γ) and interleukin-4 (IL-4) were detected by radioimmunoassay and ELISA. Anti-CD31 and anti-Ki-67 antibodies were used to evaluate neo-vascularization in xenografts and proliferation activity of ovarian follicles. Peripheral CD4+/CD8+ T cells were analyzed by flow cytometry. RESULTS: Combined treatment with cyclosporin A and FTY720 improved graft survival and reduced peripheral CD4+ and CD8+ T cell counts compared to treatment with cyclosporin A alone. Neovascularization took place in the peripheral zone of the xenograft while granulosa cells, positively stained by Ki-67, were found in early-stage follicles and stromal cells in the combined treatment groups. CONCLUSION: FTY720 in combination with cyclosporin A maintains human ovarian xenografts in these rabbit models.

Outcomes of conservative therapy for young women with early endometrial adenocarcinoma.

Four of six patients with endometrial cancer had initial response to progesterone therapy and obtained complete response; three among them had successful pregnancies with three live births. The results suggest that progesterone therapy, combined with assisted reproductive technology, provides more chance of carrying a full-term pregnancy for patients with endometrial adenocarcinoma.

Factors associated with positive margins in patients with cervical intraepithelial neoplasia grade 3 and postconization management.

OBJECTIVE: To evaluate the risk factors for positive margins in cervical intraepithelial neoplasia (CIN) grade 3 and the outcomes of postconization management. METHODS: A retrospective review of the records of 1113 women who underwent conization for CIN 3 between 2000 and 2008. RESULTS: Positive margins occurred in the following: 104 (10.7%) women with severe dysplasia versus 37 (26.2%) with carcinoma in situ; 32 (4.8%) treated with cold knife conization versus 109 (24.1%) treated with the loop electrosurgical excision procedure (LEEP); and 124 (11.6%) premenopausal versus 17 (35.4%) postmenopausal women. None of the women with severe dysplasia had invasive disease in the repeat excision specimen, whereas 3 (8.6%) women with carcinoma in situ had residual microinvasive carcinoma. CONCLUSION: LEEP, carcinoma in situ, menopausal status, and larger area of lesion are risk factors for positive margins. For women with CIN 3 and positive margins, follow-up at an interval of 6 months or repeat excision are treatment options. However, when repeat excision is technically impossible, whether simple hysterectomy or radical surgery is a rational treatment option requires further investigation.

Methotrexate induces apoptosis of human choriocarcinoma cell line JAR via a mitochondrial pathway.

OBJECTIVE: To investigate methotrexate (MTX)-induced apoptosis and the involved pathways in human choriocarcinoma cells. STUDY DESIGN: MTX-induced apoptosis of human choriocarcinoma cell line JAR was examined using a PI/Annexin V stain with flow cytometer (FCM). Mitochondrial apoptosis was detected by fluorescence microscopy, and analyzed by FCM using a MitoCapture mitochondrial apoptosis detection kit. The activities of caspase-8 and caspase-9 were quantified by microtiter plate reader at 405 nm using FLICE/Caspase-8 colorimetric assay kit and Caspase-9/Mch6 colorimetric assay kit. The changes in Bax and Bcl-2 expression were detected during apoptosis using immunocytochemistry and Western blot analysis. RESULTS: JAR cells underwent apoptosis after exposure to 0.1-2.5 microg/ml MTX for 48 h. Decreased mitochondrial membrane potential was observed both by fluorescence microscopy and FCM. The activation of caspase-9 was increased 4.35+/-0.76-fold in MTX-incubated JAR, while there was no obvious change in the activation of caspase-8. When JAR cells underwent apoptosis, the expression of Bcl-2 was decreased and the expression of Bax was increased; both were detected by immunocytochemistry assay. CONCLUSION: Methotrexate in lower concentrations induces apoptosis of human choriocarcinoma cells via mitochondrial-initiated pathways, including reduction of mitochondrial membrane potential, activation of caspase-9, and up-regulation of Bax/Bcl-2 expression.